Aqueous suspensions of tmc278

ABSTRACT

This invention concerns pharmaceutical compositions for administration via intramuscular or subcutaneous injection, comprising micro- or nanoparticles of the NNRTI compound TMC278, suspended in an aqueous pharmaceutically acceptable carrier, and the use of such pharmaceutical compositions in the treatment and prophylaxis of HIV infection.

FIELD OF THE INVENTION

This invention concerns pharmaceutical compositions for administrationvia intramuscular or subcutaneous injection, comprising micro- ornanoparticles of the NNRTI compound TMC278, suspended in an aqueouspharmaceutically acceptable carrier, and the use of such pharmaceuticalcompositions in the treatment and prophylaxis of HIV infection.

BACKGROUND OF THE INVENTION

The treatment of Human Immunodeficiency Virus (HIV) infection, known ascause of the acquired immunodeficiency syndrome (AIDS), remains a majormedical challenge. HIV is able to evade immunological pressure, to adaptto a variety of cell types and growth conditions and to developresistance against currently available drug therapies. The latterinclude nucleoside reverse transcriptase inhibitors (NRTIs),non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleotidereverse transcriptase inhibitors (NtRTIs), HIV-protease inhibitors (PIs)and the more recent fusion inhibitors.

Although effective in suppressing HIV, each of these drugs, when usedalone, is confronted with the emergence of resistant mutants. This ledto the introduction of combination therapy of several anti-HIV agentsusually having a different activity profile. In particular theintroduction of “HAART” (Highly Active Anti-Retroviral Therapy) resultedin a remarkable improvement in anti-HIV therapy, leading to a dramaticreduction in HIV-associated morbity and mortality. Current guidelinesfor antiretroviral therapy recommend such triple combination therapyregimen even for initial treatment. However, none of the currentlyavailable drug therapies is capable of completely eradicating HIV. EvenHAART can face the emergence of resistance, often due to non-adherenceand non-persistence with antiretroviral therapy. In these cases HAARTcan be made effective again by replacing one of its components by one ofanother class. If applied correctly, treatment with HAART combinationscan suppress the virus for many years, up to decades, to a level whereit no longer can cause the outbreak of AIDS.

One class of HIV drugs often used in HAART is that of the NNRTIs, anumber of which are currently on the market and several others are invarious stages of development. An NNRTI currently in development is thecompound4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]-amino]-2-pyrimidinyl]-amino]-benzonitrile,also referred to as TMC278. This compound not only shows pronouncedactivity against wild type HIV, but also against many of its mutatedvariants. The compound TMC278, its pharmacological activity as well as anumber of procedures for its preparation have been described in WO03/16306. Various conventional pharmaceutical dosage forms, includingtablets, capsules, drops, suppositories, oral solutions and injectablesolutions are exemplified therein.

Because of their pharmacokinetic properties and the need to keep plasmalevels above a minimum level, currently used anti-HIV drugs requirefrequent administration of relatively high doses. The number and/orvolume of dosage forms that need to be administered are commonlyreferred to as “pill burden”. A high pill burden is undesirable for manyreasons, such as the frequency of intake, often combined with theinconvenience of having to swallow large dosage forms, as well as theneed to store and transport a large number or volume of pills. A highpill burden increases the risk of patients not taking their entire dose,thereby failing to comply with the prescribed dosage regimen. As well asreducing the effectiveness of the treatment, this also leads to theemergence of viral resistance. The problems associated with a high pillburden are prominent in anti-HIV therapy where a patient must take alarge number of different anti-HIV agents.

Therefore, it would be desirable to provide HIV inhibitory therapy thatreduces pill burden in that it involves the administration of dosageforms of relatively small size and additionally does not requirefrequent dosing. It would be attractive to provide anti-HIV therapyinvolving the administration of dosage forms at long time intervals suchas one week or longer, or even one month or longer.

HIV can never completely be eradicated so that persons infected with HIVpose a continuous risk of infecting others. After initial infection ittakes a long time before the outbreak of the first symptoms of AIDS.People may live for years with the infection without experiencing anyeffects of it thereby being unaware of the risk of further transferringthe virus to others. Prevention of HIV transmission therefore iscrucial. Prevention currently focuses on avoiding transmission by sexualcontacts, in particular by the use of condoms in populations at risk ofbeing infected, on careful monitoring of blood samples for the presenceof HIV and on avoiding of contact with blood of potentially infectedsubjects.

Despite these measures there is always an imminent risk of individualsbeing in contact with HIV infected persons of becoming infected. This inparticular is the case for those providing medical care to infectedpatients or patients at risk of being infected such as physicians,nurses or dentists. Another group of individuals at risk are breast-fedinfants whose mother is infected or at risk of becoming infected,especially in developing countries where alternatives for breast-feedingare less obvious.

Hence there is a need for further means that provide prevention againsttransmission of HIV. There is a particular need for effective preventionmeans that are easy to apply. Providing such prevention means is anotherobject of the present invention.

It now has been found that the compound TMC278 can be formulated intomicro- or nanoparticles and that such formulations can be used as depotformulations, which may find use in the treatment of HIV infection aswell as in the prevention against transmission of HIV. Nanoparticles areknown in the prior art, having been described, for example, in EP-A-0499 299. Such particles have an average particle size in the submicronrange and consist of particles of a crystalline drug substance having asurface modifier adsorbed on their surface. Nanoparticles have been usedto formulate poorly water-soluble active ingredients.

The invention furthermore relates to the intermittent administration ofthese micro- or nanoparticle formulations at time intervals of one weekor longer that result in plasma levels that may be sufficient tosuppress the growth of HIV. This allows for a reduced number ofadministrations thereby being beneficial in terms of pill burden anddrug compliance of the patient. The micro- or nanoparticle formulationsof TMC278 of the invention therefore may be useful in the long-termtreatment of HIV infection.

The intermittent administration of micro- or nanoparticle formulationsof TMC278 at time intervals of one week or longer furthermore results inplasma levels that may be sufficient to provide prevention againsttransmission of HIV. Also in this instance, a reduced number ofadministrations is required, which again is advantageous in terms ofpill burden and drug compliance of the individual at risk of beinginfected.

SUMMARY OF THE INVENTION

The present invention is concerned with a pharmaceutical composition foradministration by intramuscular or subcutaneous injection, comprising atherapeutically effective amount of TMC278, a salt, a stereoisomer or astereoisomeric mixture thereof, in the form of a suspension of micro- ornanoparticles comprising:

-   (a) TMC278, a salt, a stereoisomer or a stereoisomeric mixture    thereof, in micro- or nanoparticle form, having a surface modifier    adsorbed to the surface thereof, and-   (b) a pharmaceutically acceptable aqueous carrier; wherein the    TMC278 active ingredient is suspended.

The invention further concerns a method of treating a subject infectedwith HIV, said method comprising the administration, by intramuscular orsubcutaneous injection, of an anti-HIV effective amount pharmaceuticalcomposition as specified above or hereinafter. Or, alternatively, theinvention concerns the use of a pharmaceutical composition as specifiedabove or hereinafter, for the manufacture of a medicament for treatingHIV infection. In one embodiment, the composition is for the long-termtreatment of HIV infection.

In another aspect, there is provided a method for the long termtreatment of a subject infected with HIV, said method comprising theadministration of an effective amount of a pharmaceutical composition asspecified above or hereinafter, for administration by intramuscular orsubcutaneous injection; wherein the composition is administered or is tobe administered intermittently at a time interval that is in the rangeof one week to one year, or one week to two years. Or, alternatively,the invention concerns the use of a pharmaceutical composition asspecified above or hereinafter, for the manufacture of a medicament forthe long term treatment of a subject infected with HIV, foradministration by intramuscular or subcutaneous injection, wherein thecomposition is administered or is to be administered intermittently at atime interval that is in the range of one week to one year, or one weekto two years.

The invention further concerns a method for the prevention of HIVinfection in a subject at risk of being infected by HIV, said methodcomprising administering an amount, effective in preventing HIVinfection, of a pharmaceutical composition as specified above or asfurther specified hereinafter, to said subject. Or alternatively, theinvention concerns the use of a pharmaceutical composition as specifiedabove or as further specified hereinafter for the manufacture of amedicament for the prevention of HIV infection in a subject at risk ofbeing infected by HIV.

In another aspect the invention relates to a method for the long termprevention of HIV infection in a subject at risk of being infected byHIV, said method comprising administering to said subject an effectiveamount of a pharmaceutical composition as specified above or as furtherspecified hereinafter, wherein the composition is administered or is tobe administered intermittently at a time interval that is in the rangeof one week to one year, or one week to two years.

The present invention furthermore relates to the use of a pharmaceuticalcomposition as specified above or as further specified hereinafter, forthe manufacture of a medicament for the long term prevention for thelong term prevention of HIV infection in a subject at risk of beinginfected by HIV, wherein the composition is administered or is to beadministered intermittently at a time interval that is in the range ofone week to one year or one week to two years.

In one embodiment the invention concerns a use or a method as specifiedherein, wherein the pharmaceutical composition is administered or is tobe administered at a time interval that is in the range of one week toone month, or in the range of one month to three months, or in the rangeof three months to six months, or in the range of six months to twelvemonths, or in the range of 12 months to 24 months.

In another embodiment the invention concerns a use or a method asspecified herein, wherein the pharmaceutical composition is administeredor is to be administered once every two weeks, or once every month, oronce every three months.

Further pharmaceutical compositions, methods of treatment or prevention,as well as uses for the manufacture of medicaments based on thesecompositions will be described hereinafter and are meant to be part ofthe present invention.

DETAILED DESCRIPTION OF THE INVENTION

The compound used in the invention is the compound TMC278 (also referredto as R278474 or rilpivirine) or4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]amino]-2-pyrimidinyl]amino]benzonitrile.

TMC278 can be used in base form or as a suitable pharmaceuticallyacceptable addition salt form, such as an acid addition salt form. Thepharmaceutically acceptable addition salts are meant to comprise thetherapeutically active non-toxic salt forms. The acid addition saltforms can be obtained by treating the base form with appropriate acidsas inorganic acids, for example, hydrohalic acids, e.g. hydrochloric,hydrobromic and the like; sulfuric acid; nitric acid; phosphoric acidand the like; or organic acids, for example, acetic, propanoic,hydroxyacetic, 2-hydroxypropanoic, 2-oxopropanoic, oxalic, malonic,succinic, maleic, fumaric, malic, tartaric,2-hydroxy-1,2,3-propanetri-carboxylic, methanesulfonic, ethanesulfonic,benzenesulfonic, 4-methylbenzene-sulfonic, cyclohexanesulfamic,2-hydroxybenzoic, 4-amino-2-hydroxybenzoic and the like acids. In oneembodiment, the TMC278 active ingredient used is the base form ofTMC278.

The term “addition salt” also comprises pharmaceutically acceptablehydrates and solvent addition forms that the compound TMC278 is able toform. Examples of such forms are e.g. hydrates, alcoholates and thelike.

TMC278 occurs in stereoisomeric forms, more in particular as E- andZ-isomeric forms. Both isomers may be used in the present invention.Whenever reference is made herein to TMC278, the E- or the Z-form aswell as any mixture of both forms are meant to be included. A preferredform of TMC278 for use in the invention is the E-isomer, i.e.(E)-4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]-amino]-2-pyrimidinyl]-amino]-benzonitrile,which may be referred to as E-TMC278. The Z-isomer of TMC278, i.e.(Z)-4-[[4-[[4-(2-cyanoethenyl)-2,6-dimethylphenyl]-amino]-2-pyrimidinyl]-amino]-benzonitrile,which may be referred to as Z-TMC278), can also be used.

Whenever reference is made herein to the E-form of TMC278 (i.e.E-TMC278), the pure E-isomer or any isomeric mixture of the E- and theZ-forms wherein the E-form is predominantly present is meant to becomprised, i.e. an isomeric mixture containing more than 50% or inparticular more than 80% of the E-form, or even more than 90% of theE-form. Of particular interest is the E-form substantially free of theZ-form. Substantially free in this context refers to E-Z-mixtures withno or almost no Z-form, e.g. isomeric mixtures containing as much as90%, in particular 95% or even 98% or 99% of the E-form. Equally,whenever reference is made herein to the Z-form of TMC278 (i.e.Z-TMC278), the pure Z-isomer or any isomeric mixture of the Z- and theE-forms wherein the Z-form is predominantly present is meant to becomprised, i.e. an isomeric mixture containing more than 50% or inparticular more than 80% of the Z-form, or even more than 90% of theZ-form. The Z-form substantially free of the E-form can also be used.Substantially free in this context refers to E-Z-mixtures with no oralmost no E-form, e.g. isomeric mixtures containing as much as 90%, inparticular 95% or even 98% or 99% of the Z-form. In one embodiment, theTMC278 active ingredient used is the E-form of TMC278, in particular theE-form of TMC278 base.

Also meant to be included for use in this invention are salts of thestereoisomeric forms of TMC278, in particular the salts mentioned aboveof Z-TMC278 or of E-TMC278.

Whenever used herein, the term “TMC278” refers to as well the base formas any pharmaceutically acceptable acid-addition salt thereof, and alsoto the stereoisomeric forms of TMC278 as well as any pharmaceuticallyacceptable acid-addition salt of said stereoisomeric forms. Inparticular, the term “TMC278” refers to the E-isomer of TMC278 as wellas its pharmaceutically acceptable acid-addition salts. The term“TMC278” may also refer to the base form of the E-isomer of TMC278.

It has been found that the physico-chemical properties of TMC278 allowfor the manufacture of micro- or nanoparticle suspensions that haveunique pharmacokinetic properties in that they can be used for thelong-term treatment of HIV infection as well as in the long-termprevention of HIV infection and to this purpose only a limited number ofdrug administrations is required. This is beneficial in terms ofpill-burden as well as patient compliance with the prescribed doseregimen.

As used herein the term “treatment of HIV infection” relates to thetreatment of a subject being infected with HIV. The term “treatment ofHIV infection” also relates to the treatment of diseases associated withHIV infection, for example AIDS, or other conditions associated with HIVinfection including thrombocytopaenia, Kaposi's sarcoma and infection ofthe central nervous system characterized by progressive demyelination,resulting in dementia and symptoms such as, progressive dysarthria,ataxia and disorientation, and further conditions where HIV infectionhas also been associated with, such as peripheral neuropathy,progressive generalized lymphadenopathy (PGL), and AIDS-related complex(ARC).

The term “prevention of HIV infection” relates to the prevention oravoidance of a subject becoming infected with HIV. The source ofinfection can be various, a material containing HIV, in particular abody fluid that contains HIV such as blood or sperm, or another subjectwho is infected with HIV. Prevention of HIV infection relates to theprevention of the transmission of the virus from the material containingHIV or from the HIV infected individual to an uninfected person, orrelates to the prevention of the virus from entering the body from anuninfected person. Transmission of the HIV virus can be by any knowncause of HIV transfer such as by sexual transmission or by contact withblood of an infected subject, e.g. medical staff providing care toinfected subjects. Transfer of HIV can also occur by contact with HIVinfected blood, e.g. when handling blood samples or with bloodtransfusion. It can also be by contact with infected cells, e.g. whencarrying out laboratory experiments with HIV infected cells.

The terms “treatment of HIV infection”, “anti-HIV therapy”, as well assimilar terms, refer to a treatment by which the viral load of HIV(represented as the number of copies of viral RNA in a specified volumeof serum) is reduced. The more effective the treatment, the lower theviral load. Preferably the viral load should be reduced to as low levelsas possible, e.g. below about 200 copies/ml, in particular below about100 copies/ml, more in particular below 50 copies/ml, if possible belowthe detection limit of the virus. Reductions of viral load of one, twoor even three orders of magnitude (e.g. a reduction in the order ofabout 10 to about 10², or more, such as about 10³) are an indication ofthe effectiveness of the treatment. Another parameter to measureeffectiveness of anti-HIV treatment is the CD4 count, which in normaladults ranges from 500 to 1500 cells per μl. Lowered CD4 counts are anindication of HIV infection and once below about 200 cells per μl, AIDSmay develop. An increase of CD4 count, e.g. with about 50, 100, 200 ormore cells per μl, is also an indication of the effectiveness ofanti-HIV treatment. The CD4 count in particular should be increased to alevel above about 200 cells per μl, or above about 350 cells per μl.Viral load or CD4 count, or both, can be used to diagnose the degree ofHIV infection.

The terms “effective treatment of HIV” and similar terms refer to thattreatment that lowers the viral load, or increases CD4 count, or both,as described above. The terms “effective prevention of HIV” and similarterms refer to that situation where there is a decrease in the relativenumber of newly infected subjects in a population in contact with asource of HIV infection such as a material containing HIV, or a HIVinfected subject. Effective prevention can be measured, for example, bymeasuring in a mixed population of HIV infected and non-infectedindividuals, if there is a decrease of the relative number of newlyinfected individuals, when comparing non-infected individuals treatedwith a pharmaceutical composition of the invention, and non-treatednon-infected individuals. This decrease can be measured by statisticalanalysis of the numbers of infected and non-infected individuals in agiven population over time.

The terms “therapeutically effective amount”, “an amount, effective inpreventing HIV infection”, and similar terms, refer to amounts of theactive ingredient TMC278 that result in efficacious blood plasma levels.With “efficacious blood plasma levels” is meant those blood plasmalevels of the HIV inhibitor TMC278 that provide effective treatment oreffective prevention of HIV infection.

The term “subject” in particular relates to a human being.

The term “micro- or nanoparticles” refers to particles in the micrometeror nanometer range. The size of the particles should be below a maximumsize above which administration by subcutaneous or intramuscularinjection becomes impaired or even no longer is possible. Said maximumsize depends for example on the limitations imposed by the needlediameter or by adverse reactions of the body to large particles, orboth. In one embodiment, the pharmaceutical compositions of theinvention comprise TMC278 in nanoparticle form.

The average effective particle size of the micro- or nanoparticles ofthe present invention may be below about 50 μm, or below about 20 μm, orbelow about 10 μm, or below about 1000 nm, or below about 500 nm, orbelow about 400 nm, or below about 300 nm, or below about 200 nm. Thelower limit of the average effective particle size may be low, e.g. aslow as about 100 nm or as low as about 50 nm. In one embodiment, theaverage effective particle size is in the range of about 50 nm to about50 μm, or about 50 nm to about 20 μm, or about 50 nm to about 10 μm, orabout 50 nm to about 1000 nm, about 50 nm to about 500 nm, or about 50nm to about 400 nm, or about 50 nm to about 300 nm, or about 50 nm toabout 250 nm, or about 100 nm to about 250 nm, or about 150 nm to about220 nm, or 100 to 200 nm, or about 150 nm to about 200 nm, e.g. about130 nm, or about 150 nm.

As used herein, the term average effective particle size has itsconventional meaning as known to the person skilled in the art and canbe measured by art-known particle size measuring techniques such as, forexample, sedimentation field flow fractionation, photon correlationspectroscopy, laser diffraction or disk centrifugation. The averageeffective particle sizes mentioned herein may be related to volumedistributions of the particles. In that instance, by “an effectiveaverage particle size of less than about 50 μm” it is meant that atleast 50% of the volume of the particles has a particle size of lessthan the effective average of 50 μm, and the same applies to the othereffective particle sizes mentioned. In a similar manner, the averageeffective particle sizes may be related to weight distributions of theparticles but usually this will result in the same or about the samevalue for the average effective particle size.

The pharmaceutical compositions of the present invention provide releaseof the active ingredient TMC278 over a prolonged period of time andtherefore they can also be referred to as sustained or delayed releasecompositions. After administration, the compositions of the inventionstay in the body and steadily release TMC278, keeping such levels ofthis active ingredient in the patient's system for a prolonged period oftime, thereby providing, during said period, anti-HIV therapy orprevention of HIV infection. Because of the fact that the pharmaceuticalcompositions of the invention stay in the body and steadily releaseTMC278, they can be referred to as pharmaceutical compositions suitableas depot formulations.

As used herein with the term “prolonged period of time”, there is meanta term (or time period) that may be in the range of one week up to oneyear or up to two years, or a term in the range of one to two weeks, ortwo to three weeks, or three to four weeks, or a term in the range ofone to two months, or two to three months, or three to four months, orthree to six months, or six months to 12 months, or 12 months to 24months, or a term that is in the range of several days, e.g. 7, 10 or 12days, or several weeks, e.g. 2, 3 or 4 weeks, or one month, or severalmonths, e.g. 2, 3, 4, 5 or six months or even longer, e.g. 7, 8, 9 or 12months.

The pharmaceutical compositions of this invention may be applied in thelong-term treatment or the long-term prevention of HIV infection, orwith other words they may be used in the treatment of HIV infection, orin the prevention of HIV infection, during a prolonged period of time.The compositions of the invention are effective in anti-HIV therapy orin the prevention of HIV infection for a prolonged period of time, forexample for at least about one week or longer, or for about 1 month orlonger. By the expression “effective for at least about one week orlonger”, one means that the plasma level of the active ingredient,TMC278, should be above a threshold value. In case of therapeuticapplication said threshold value is the lowest plasma level at whichTMC278 provides effective treatment of HIV infection. In case ofapplication in the prevention of HIV infection said threshold value isthe lowest plasma level at which TMC278 is effective in preventingtransmission of HIV infection.

With “long term” for example as used in relation to “long termprevention of HIV infection” or “long term treatment of HIV infection”,or similar terminology, there are meant terms that may be in the rangeof one week up to one year or up to two years, or longer, such as fiveor 10 years. In particular in the case of treatment of HIV infection,such terms will be long, in the order of one to several years. Suchterms may also be relatively short, in particular in the case ofprevention. Shorter terms are those of several days, e.g. 7, 10 or 12days, or several weeks, e.g. 2, 3 or 4 weeks, or one month, or severalmonths, e.g. 2, 3, 4, 5 or six months or even longer, e.g. 7, 8, 9 or 12months. In one embodiment the methods and uses in accordance with thepresent invention are for the prevention of HIV infection during onemonth, or several months, e.g. 2, 3, 4, 5 or six months or even longer,e.g. 7, 8, 9 or 12 months.

The pharmaceutical compositions of the present invention can beadministered at various time intervals. When used in the prevention ofHIV infection, the pharmaceutical compositions of this invention can beadministered only once or a limited number of times such as twice,three, four, five or six times, or more. This may be recommendable whereprevention is required during a limited period of time, such as theperiod during which there is a risk of infection.

The pharmaceutical compositions of the present invention can beadministered at the time intervals mentioned above, such as at a timeinterval that is in the range of one week to one month, or in the rangeof one month to three months, or in the range of three months to sixmonths, or in the range of six months to twelve months. In oneembodiment, the pharmaceutical composition can be administered onceevery two weeks, or once every month, or once every three months. Inanother embodiment the time interval is in the range of one to twoweeks, or two to three weeks, or three to four weeks, or the timeinterval is in the range of one to two months, or two to three months,or three to four months, or three to six months, or six months to 12months, or 12 months to 24 months. The time interval may be at least oneweek, but may also be several weeks, e.g. 2, 3, 4, 5 or 6 weeks, or attime intervals of one month, or of several months, e.g. 2, 3, 4, 5 or 6months or even longer, e.g. 7, 8, 9 or 12 months. In one embodiment, thepharmaceutical compositions of the present invention are administered ata time interval of one, two or three months. These longer periodsbetween each administration of the pharmaceutical compositions of theinvention provide further improvements in terms of pill burden andcompliance. To further improve compliance, patients can be instructed totake their medication at a certain day of the week, where thecomposition is administered on a weekly schedule, or at a certain day ofthe month in case of a monthly schedule.

The length of the time intervals between each administration of acomposition of the present invention may vary. For example said timeintervals may be selected in function of the blood plasma levels. Theintervals may be shorter where the blood plasma levels of TMC278 aredeemed too low, e.g. when these approach the minimum blood plasma levelspecified hereinafter. The intervals may be longer where the bloodplasma levels of TMC278 are deemed too high. In one embodiment, thecompositions of the invention are administered at equal time intervals.The compositions may be administered without any interjacent additionaladministrations, or with other words, the compositions may beadministered at particular points in time separated from one another bya time period of varying or equal length, e.g. a time period of at leastone week, or any other time period specified herein, during which nofurther TMC278 is administered. Having time intervals of the same lengthhas the advantage that the administration schedule is simple, e.g.administration takes place at the same day in the week, or the same dayin the month. Such administration schedule therefore involves limited“pill burden” thereby contributing beneficially to the patient'scompliance to the prescribed dosing regimen.

The concentration (or “C”) of TMC278 in the blood plasma of a subjecttreated therewith is generally expressed as mass per unit volume,typically nanograms per milliliter (ng/ml). For convenience, thisconcentration may be referred to herein as “blood plasma drugconcentration” or “blood plasma concentration”.

The dose (or amount) of TMC278 administered, depends on the amount ofTMC278 in the pharmaceutical compositions of the invention, or on theamount of a given composition that is administered. Where higher bloodplasma levels are desired, either or both of a composition of higherTMC278 concentration, or more of a given composition, may beadministered. This applies vice versa if lower plasma levels aredesired. Also a combination of varying time intervals and varying dosingmay be selected to attain certain desired blood plasma levels.

The dose (or amount) of TMC278 administered also depends on thefrequency of the administrations (i.e. the time interval between eachadministration). Usually, the dose will be higher where administrationsare less frequent. All these parameters can be used to direct the bloodplasma levels to desired values

The dosing regimen also depends on whether prevention or treatment ofHIV infection is envisaged. In case of therapy, the dose of TMC278administered or the frequency of dosing, or both, are selected so thatthe blood plasma concentration of TMC278 is kept above a minimum bloodplasma level. The term “minimum blood plasma level” (or C_(min)) in thiscontext refers to the blood plasma level of TMC278 that provideseffective treatment of HIV. In particular, the blood plasma level ofTMC278 is kept at a level above a minimum blood plasma level of about 10ng/ml, or above about 15 ng/ml, or above about 20 ng/ml, or above about40 ng/ml. The blood plasma level of TMC278 may be kept above a minimumblood plasma level that is higher, for example above about 50 ng/ml, orabove about 90 ng/ml, or above about 270 ng/ml, or above about 540 ng/mlIn one embodiment, the blood plasma level of TMC278 is kept above alevel of about 13.5 ng/ml, or is kept above a level of about 20 ng/ml.Or the blood plasma level of TMC278 may be kept within certain ranges,in particular ranges starting from a minimum blood plasma level selectedfrom those mentioned above and ending at a higher blood plasma levelsselected from those mentioned above and selected from 500 ng/ml and 1000ng/ml (e.g. from 10 to 15, 10 to 20, 10 to 40, etc., or from 15 to 20,or 15 to 40, or 15 to 90, etc., or 20 to 40, 20 to 90, or 20 to 270,etc., or 40 to 90, 40 to 270, or 40-540, etc., each time from about theindicated value in ng/ml to about the indicated value in ng/ml). In oneembodiment said range is from about 10 to about 20, from about 20 toabout 90, from 90 to 270, from 270 to 540, from 540 to 1000, each timefrom about the indicated value in ng/ml to about the indicated value inng/ml.

The plasma levels of TMC278 should be kept above the above-mentionedminimum blood plasma levels because at lower levels the virus may nolonger be sufficiently suppressed so that it can multiply with theadditional risk of the emergence of mutations.

In the instance of HIV prevention, the term “minimum blood plasma level”(or C_(min)) refers to the lowest blood plasma level of TMC278 thatprovides effective prevention of HIV infection. In the case oftransmission of HIV from a material containing HIV or from a subjectinfected by HIV to a subject not infected by HIV, this is the lowestblood plasma level that is effective in inhibiting said transmission.

In particular, in the instance of HIV prevention, the blood plasma levelof TMC278 can be kept at a level above a minimum blood plasma levelmentioned above in relation to therapy. However in prevention the bloodplasma level of TMC278 can be kept at a lower level, for example at alevel above about 4 ng/ml, or about 5 ng/ml, or about 8 ng/ml. The bloodplasma levels of TMC278 should preferably be kept above these minimumblood plasma levels because at lower levels the drug may no longer beeffective thereby increasing the risk of transmission of HIV infection.Plasma levels of TMC278 may be kept at somewhat higher levels to have asafety margin. Such higher levels start from about 50 ng/ml or more. Theblood plasma level of TMC278 can be kept at a level that is in theranges mentioned above in relation to therapy, but where the lowerlimits include the blood plasma levels of about 4 ng/ml, or about 5ng/ml, or about 8 ng/ml.

An advantage of TMC278 is that it can be used up to relatively highblood plasma levels without any significant side effects. The plasmaconcentrations of TMC278 may reach be relatively high levels, but aswith any drug should not exceed a maximum plasma level (or C_(max)),which is the blood plasma level where TMC278 causes significant sideeffects. As used herein, the term “significant side effects” means thatthe side effects are present in a relevant patient population to anextend that the side effects affect the patients' normal functioning.The C_(max) for TMC278 can be determined from the extrapolation of testdata in cellular assays or from the evaluation of clinical testing andpreferably should not exceed a value of about 500 ng/ml or 1000 ng/ml.In an embodiment, the amount and the frequency of administrations ofTMC278 to be administered are selected such that the blood plasmaconcentrations are kept during a long term at a level comprised betweena maximum plasma level (or C_(max) as specified above) and a minimumblood plasma level (or C_(min) as specified above).

In certain instances it may be desirable to keep the plasma levels ofTMC278 at relatively low levels, e.g. as close as possible to theminimum blood plasma levels specified herein. This will allow reducingthe frequency of the administrations and/or the quantity of TMC278administered with each administration. It will also allow avoidingundesirable side effects, which will contribute to the acceptance of thedosage forms in most of the targeted population groups who are healthypeople at risk of being infected and therefore are less inclined totolerate side effects. The plasma levels of TMC278 may be kept atrelatively low levels in the instance of prevention. One embodimentconcerns uses or methods for prevention of HIV infection, as specifiedabove or hereinafter, wherein the minimum blood plasma level of TMC278is as specified herein and the maximum blood plasma level is about equalto the lowest blood plasma level that causes the RT inhibitor to acttherapeutically, also as specified herein.

In other embodiments, the blood plasma level of TMC278 is kept at alevel below a lower maximum blood plasma level of about 10 ng/ml, morein particular about 15 ng/ml, further in particular about 20 ng/ml,still more in particular about 40 ng/ml. In a particular embodiment, theblood plasma level of TMC278 is kept below a level of about 13.5 ng/ml.In one embodiment, the plasma level of TMC 278 is kept in an interval ofthe lower maximum blood level specified above, and the minimum bloodplasma levels mentioned in relation to prevention. For example the bloodplasma levels of TMC278 are kept below about 10 ng/ml and above aminimum level of about 4 ng/ml.

In other instances it may be desirable to keep the plasma levels ofTMC278 at relatively higher levels, for example where there is a highrisk of infection and more frequent and/or higher doses are not anissue. In these instances the minimum blood plasma level may be equal tothe lowest blood plasma level of TMC278 that provides effectivetreatment of HIV, such as the specific levels mentioned herein.

In the instance of prevention, the dose to be administered should becalculated on a basis of about 0.2 mg/day to about 50 mg/day, or 0.5mg/day to about 50 mg/day, or of about 1 mg/day to about 10 mg/day, orabout 2 mg/day to about 5 mg/day, e.g. about 3 mg/day. This correspondsto a weekly dose of about 1.5 mg to about 350 mg, in particular of about3.5 mg to about 350 mg, in particular of about 7 mg to about 70 mg, orabout 14 mg to about 35 mg, e.g. about 35 mg, or to a monthly dose offrom 6 mg to about 3000 mg, in particular about 15 mg to about 1,500 mg,more in particular of about 30 mg to about 300 mg, or about 60 mg toabout 150 mg, e.g. about 150 mg. Doses for other dosing regimens canreadily be calculated by multiplying the daily dose with the number ofdays between each administration.

In the instance of therapy, the dose to be administered should besomewhat higher and should be calculated on a basis of about 1 mg/day toabout 150 mg/day, or of about 2 mg/day to about 100 mg/day, or of about5 mg/day to about 50 mg/day, or about 10 mg/day to about 25 mg/day, e.g.about 15 mg/day. The corresponding weekly or monthly doses can becalculated as set forth above. For applications in prevention, the dosesmay be lower although the same dosing as for therapeutic applicationsmay be used.

It has been found that, once administered, the blood plasma levels ofTMC278 are more or less stable, i.e. they fluctuate within limitedmargins. The blood plasma levels have been found to approach more orless a steady state mode or to approximate more or less a zero orderrelease rate during a prolonged period of time. By “steady state” ismeant the condition in which the amount of drug present in the bloodplasma of a subject stays at more or less the same level over aprolonged period of time. The plasma levels of TMC278 generally do notshow any drops below the minimum plasma level at which the drug iseffective. The term “stays at more or less the same level” does notexclude that there can be small fluctuations of the plasmaconcentrations within an acceptable range, e.g. fluctuations within arange of about +/−30%, or about +/−20%, or about +/−10%, or about+/−10%.

In some instances there may be an initial plasma concentration peakafter administration, after which the plasma levels achieve a“steady-state”, as mentioned hereinafter.

The compositions of the invention show good local tolerance and ease ofadministration. Good local tolerance relates to minimal irritation andinflammation at the site of injection; ease of administration refers tothe size of needle and length of time required to administer a dose of aparticular drug formulation. In addition, the compositions of theinvention show good stability and have an acceptable shelf life.

The micro- or nanoparticles of the present invention have a surfacemodifier adsorbed on the surface thereof. The function of the surfacemodifier is to act as a wetting agent as well as a stabilizer of thecolloidial suspension.

In one embodiment, the micro- or nanoparticles in the compositions ofthe invention mainly comprise crystalline TMC278 or a salt thereof, andthe surface modifier, the combined amount of which may at least compriseabout 50%, or at least about 80%, or at least about 90%, or at leastabout 95%, or at least about 99% of the micro- or nano particles.

In a further aspect, the present invention is concerned with apharmaceutical composition for administration by intramuscular orsubcutaneous injection, comprising a therapeutically effective amount ofTMC278, or a stereoisomer or a stereoisomeric mixture thereof, in theform of a suspension of particles consisting essentially of:

-   (1) TMC278, or a stereoisomer or a stereoisomeric mixture thereof in    micro- or nanoparticle form, having a surface modifier adsorbed to    the surface thereof, and-   (2) a pharmaceutically acceptable aqueous carrier; wherein the    active ingredient is suspended.

Suitable surface modifiers can be selected from known organic andinorganic pharmaceutical excipients, including various polymers, lowmolecular weight oligomers, natural products and surfactants. Particularsurface modifiers include nonionic and anionic surfactants.Representative examples of surface modifiers include gelatin, casein,lecithin, salts of negatively charged phospholipids or the acid formthereof (such as phosphatidyl glycerol, phosphatidyl inosite,phosphatidyl serine, phosphatic acid, and their salts such as alkalimetal salts, e.g. their sodium salts, for example egg phosphatidylglycerol sodium, such as the product available under the tradenameLipoid™ EPG), gum acacia, stearic acid, benzalkonium chloride,polyoxyethylene alkyl ethers, e.g., macrogol ethers such as cetomacrogol1000, polyoxyethylene castor oil derivatives; polyoxyethylene stearates,colloidal silicon dioxide, sodium dodecylsulfate, carboxymethylcellulosesodium, bile salts such as sodium taurocholate, sodiumdesoxytaurocholate, sodium desoxycholate; methylcellulose,hydroxyethylcellulose, hydroxypropylcellulose,hydroxypropyl-methylcellulose, magnesium aluminate silicate, polyvinylalcohol (PVA), poloxamers, such as Pluronic™ F68, F108 and F127 whichare block copolymers of ethylene oxide and propylene oxide; tyloxapol;Vitamin E-TGPS (α-tocopheryl polyethylene glycol succinate, inparticular α-tocopheryl polyethylene glycol 1000 succinate);poloxamines, such as Tetronic™ 908 (T908) which is a tetrafunctionalblock copolymer derived from sequential addition of ethylene oxide andpropylene oxide to ethylenediamine; dextran; lecithin; dioctyl ester ofsodium sulfosuccinic acid such as the products sold under the tradenameAerosol OT™ (AOT); sodium lauryl sulfate (Duponol™ P); alkyl arylpolyether sulfonate available under the tradename Triton™ X-200;polyoxyethylene sorbitan fatty acid esters (Tweens™ 20, 40, 60 and 80);sorbitan esters of fatty acids (Span™ 20, 40, 60 and 80 or Arlacel™ 20,40, 60 and 80); polyethylene glycols (such as those sold under thetradename Carbowax™ 3550 and 934); sucrose stearate and sucrosedistearate mixtures such as the product available under the tradenameCrodesta™ 110 or Crodesta™ SL-40; hexyldecyl trimethyl ammonium chloride(CTAC); polyvinylpyrrolidone (PVP). If desired, two or more surfacemodifiers can be used in combination.

Particular surface modifiers are selected from poloxamers, α-tocopherylpolyethylene glycol succinates, polyoxyethylene sorbitan fatty acidesters, and salts of negatively charged phospholipids or the acid formthereof. More in particular the surface modifiers are selected fromPluronic™ F108, Vitamin E TGPS, Tween™ 80, and Lipoid™ EPG. One or moreof these surface modifiers may be used. Pluronic™ F108 corresponds topoloxamer 338 and is the polyoxyethylene, polyoxypropylene blockcopolymer that conforms generally to the formulaHO—[CH₂CH₂O]_(x)—[CH(CH₃)CH₂O]_(y)—[CH₂CH₂O]_(z)—H in which the averagevalues of x, y and z are respectively 128, 54 and 128. Other commercialnames of poloxamer 338 are Hodag Nonionic™ 1108-F and Synperonic™PE/F108. In one embodiment, the surface modifier comprises a combinationof a polyoxyethylene sorbitan fatty acid ester and a phosphatidylglycerol salt (in particular egg phosphatidyl glycerol sodium).

The optimal relative amount of TMC278 in relation to the surfacemodifier depends on the surface modifier selected, the specific surfacearea of the TMC278 suspension which is determined by the averageeffective particle size and the TMC278 concentration, the criticalmicelle concentration of the surface modifier if it forms micelles, etc.The relative amount (w/w) of TMC278 to the surface modifier preferablyis in the range of 1:2 to about 20:1, in particular in the range of 1:1to about 10:1, e.g. about 4:1.

The particles of this invention can be prepared by means ofmicronization/particle size reduction/nanonization by mechanical meansand by controlled precipitation from a supersaturated solution, or byusing supercritical fluids such as in the GAS technique (“gasanti-solvent”), or any combination of such techniques. In one embodimenta method is used comprising the steps of dispersing TMC278 in a liquiddispersion medium and applying mechanical means in the presence ofgrinding media to reduce the particle size of TMC278 to an averageeffective particle size of less than about 50 μm, in particular lessthan about 1,000 nm. The particles can be reduced in size in thepresence of a surface modifier.

A general procedure for preparing the particles of this inventioncomprises

-   (a) obtaining TMC278 in micronized form;-   (b) adding the micronized TMC278 to a liquid medium to form a    premix/predispersion; and-   (c) subjecting the premix to mechanical means in the presence of a    grinding medium to reduce the average effective particle size.

TMC278 in micronized form is prepared using techniques known in the art.It is preferred that the average effective particle size of the TMC278active agent in the predispersion be less than about 100 μm asdetermined by sieve analysis. Where the average effective particle sizeof the micronized TMC278 is greater than about 100 μm, it is preferredthat the particles of the TMC278 compound be reduced in size to lessthan 100 μm.

The micronized TMC278 can then be added to a liquid medium in which itis essentially insoluble to form a predispersion. The concentration ofTMC278 in the liquid medium (weight by weight percentage) can varywidely and depends on the selected surface modifier and other factors.Suitable concentrations of TMC278 in compositions vary between about0.1% to about 60%, or between about 1% to about 60%, or between about10% to about 50%, or between about 10% to about 30%, e.g. about 10%, 20%or 30% (each % in this paragraph relating to w/v).

The premix can be used directly by subjecting it to mechanical means toreduce the effective average effective particle size in the dispersionto less than 2,000 nm. It is preferred that the premix be used directlywhen a ball mill is used for attrition. Alternatively, TMC278 and,optionally, the surface modifier, can be dispersed in the liquid mediumusing suitable agitation such as, for example, a roller mill, until ahomogeneous dispersion is achieved.

The mechanical means applied to reduce the effective average effectiveparticle size of TMC278 conveniently can take the form of a dispersionmill. Suitable dispersion mills include a ball mill, anattritor/attrition mill, a vibratory mill, a planetary mill, mediamills, such as a sand mill and a bead mill. A media mill is preferreddue to the relatively shorter milling time required to provide thedesired reduction in particle size. The beads preferably are ZrO₂ beads.

The grinding media for the particle size reduction step can be selectedfrom rigid media preferably spherical or particulate in form having anaverage size less than 3 mm and, more preferably, less than 1 mm (as lowas 200 μm beads). Such media desirably can provide the particles of theinvention with shorter processing times and impart less wear to themilling equipment. Examples of grinding media are ZrO₂ such as 95% ZrO₂stabilized with magnesia or stabilized with yttrium, zirconium silicate,glass grinding media, polymeric beads, stainless steel, titania, aluminaand the like. Preferred grinding media have a density greater than 2.5g/cm³ and include 95% ZrO₂ stabilized with magnesia and polymeric beads.

The attrition time can vary widely and depends primarily upon theparticular mechanical means and processing conditions selected. Forrolling mills, processing times of up to two days or longer may berequired.

The particles should be reduced in size at a temperature that does notsignificantly degrade the TMC278 compound. Processing temperatures ofless than 30 to 40° C. are ordinarily preferred. If desired, theprocessing equipment may be cooled with conventional cooling equipment.The method is conveniently carried out under conditions of ambienttemperature and at processing pressures, which are safe and effectivefor the milling process.

The pharmaceutical compositions according to the present inventioncontain an aqueous carrier that preferably is pharmaceuticallyacceptable. Said aqueous carrier comprises sterile water optionally inadmixture with other pharmaceutically acceptable ingredients. The lattercomprise any ingredients for use in injectable formulations. Theseingredients may be selected from one or more of a suspending agent, abuffer, a pH adjusting agent, a preservative, an isotonizing agent, andthe like ingredients. In one embodiment, said ingredients are selectedfrom one or more of a suspending agent, a buffer, a pH adjusting agent,and optionally, a preservative and an isotonizing agent. Particularingredients may function as two or more of these agents simultaneously,e.g. behave like a preservative and a buffer, or behave like a bufferand an isotonizing agent.

Suitable buffering agents and pH adjusting agents should be used inamount sufficient to render the dispersion neutral to very slightlybasic (up to pH 8.5), preferably in the pH range of 7 to 7.5. Particularbuffers are the salts of week acids. Buffering and pH adjusting agentsthat can be added may be selected from tartaric acid, maleic acid,glycine, sodium lactate/lactic acid, ascorbic acid, sodiumcitrates/citric acid, sodium acetate/acetic acid, sodiumbicarbonate/carbonic acid, sodium succinate/succinic acid, sodiumbenzoate/benzoic acid, sodium phosphates,tris(hydroxymethyl)aminomethane, sodium bicarbonate/sodium carbonate,ammonium hydroxide, benzene sulfonic acid, benzoate sodium/acid,diethanolamine, glucono delta lactone, hydrochloric acid, hydrogenbromide, lysine, methanesulfonic acid, monoethanolamine, sodiumhydroxide, tromethamine, gluconic, glyceric, gluratic, glutamic,ethylene diamine tetraacetic (EDTA), triethanolamine, including mixturesthereof.

Preservatives comprise antimicrobials and anti-oxidants which can beselected from the group consisting of benzoic acid, benzyl alcohol,butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),chlorbutol, a gallate, a hydroxybenzoate, EDTA, phenol, chlorocresol,metacresol, benzethonium chloride, myristyl-γ-piccolinium chloride,phenylmercuric acetate and thimerosal. Radical scavengers include BHA,BHT, Vitamin E and ascorbyl palmitate, and mixtures thereof. Oxygenscavengers include sodium ascorbate, sodium sulfite, L-cysteine,acetylcysteine, methionine, thioglycerol, acetone sodium bisulfite,isoacorbic acid, hydroxypropyl cyclodextrin. Chelating agents includesodium citrate, sodium EDTA and malic acid.

An isotonizing agent or isotonifier may be present to ensure isotonicityof the pharmaceutical compositions of the present invention, andincludes sugars such as glucose, dextrose, sucrose, fructose, trehalose,lactose; polyhydric sugar alcohols, preferably trihydric or higher sugaralcohols, such as glycerin, erythritol, arabitol, xylitol, sorbitol andmannitol. Alternatively, sodium chloride, sodium sulfate, or otherappropriate inorganic salts may be used to render the solutionsisotonic. These isotonifiers can be used alone or in combination. Thesuspensions conveniently comprise from 0 to 10% (w/v), in particular 0to 6% of isotonizing agent. Of interest are nonionic isotonifiers, e.g.glucose, as electrolytes may affect colloidal stability.

A desirable feature for a pharmaceutical composition of the inventionrelates to the ease of administration. The viscosity of thepharmaceutical compositions of the invention should be sufficiently lowto allow administration by injection. In particular they should bedesigned so that they can be taken up easily in a syringe (e.g. from avial), injected through a fine needle (e.g. a 20 G 1½, 21 G 1½, 22 G 2or 22 G 1¼ needle) in not too long a time span. In one embodiment theviscosity of the compositions of the invention is below about 75 mPa·s,or below 60 mPa·s. Aqueous suspensions of such viscosity or lowerusually meet the above-mentioned criteria.

Ideally, the aqueous suspensions according to the present invention willcomprise as much TMC278 as can be tolerated so as to keep the injectedvolume to a minimum, in particular from 3 to 40% (w/v), or from 3 to 30%(w/v), or from 3 to 20% (w/v), or from 10 to 30% (w/v), of TMC278. Inone embodiment the aqueous suspensions of the invention contain about10% (w/v) of TMC278, or about 10% (w/v) of TMC278, or about 30% (w/v) ofTMC278.

In one embodiment, the aqueous suspensions may comprise by weight, basedon the total volume of the composition:

-   (a) from 3% to 50% (w/v), or from 10% to 40% (w/v), or from 10% to    30% (w/v), of TMC278;-   (b) from 0.5% to 10%, or from 0.5% to 2% (w/v) of a wetting agent;-   (c) from 0% to 10%, or from 0% to 5%, or from 0% to 2%, or from 0%    to 1% of one or more buffering agents;-   (d) from 0% to 10%, or from 0% to 6% (w/v) of a isotonizing agent-   (e) from 0% to 2% (w/v) preservatives; and-   (f) water for injection q.s. ad 100%.

To the suspensions may optionally be added an amount of acid or base tobring the pH to a value of about pH 7. Suitable acids or bases are anyof those that are physiologically acceptable, e.g. HCl, HBr, sulfuricacid, alkali metal hydroxides such as NaOH.

The administration of TMC278 as in the present invention may suffice totreat HIV infection although in a number of cases it may berecommendable to co-administer other HIV inhibitors. The latterpreferably include HIV inhibitors of other classes, in particular thoseselected from NRTIs, PIs and fusion inhibitors. In one embodiment, theother HIV inhibitor that is co-administered is a PI inhibitor. HIVinhibitors that may be co-administered by preference are those used inHAART combinations comprising an NNRTI. For example two further NRTIs oran NRTI and a PI may be co-administered. Such co-administration may beby oral administration or parenterally.

In certain instances, the treatment of HIV infection may be limited toonly the administration of a composition of TMC278 in accordance withthis invention, i.e. as monotherapy without co-administration of furtherHIV inhibitors. This option may be recommended, for example, where theviral load is relatively low, for example where the viral load(represented as the number of copies of viral RNA in a specified volumeof serum) is below about 200 copies/ml, in particular below about 100copies/ml, more in particular below 50 copies/ml, specifically below thedetection limit of the virus. In one embodiment, this type ofmonotherapy is applied after initial treatment with a combination of HIVdrugs, in particular with any of the HAART combinations during a certainperiod of time until the viral load in blood plasma reaches the aforementioned low viral level.

In a further aspect the present invention relates to the use of apharmaceutical composition comprising an anti-virally effective amountof TMC278 or a pharmaceutically acceptable acid-addition salt thereof,in accordance with the present invention, for the manufacture of amedicament for maintenance therapy of a subject being infected with HIV,wherein the composition is administered or is to be administeredintermittently at a time interval that is in the range of one week toone year, or one week to two years.

Thus in a further aspect, the present invention provides a method forthe long term treatment of a patient being infected with HIV, saidmethod comprising

-   (i) the treatment of said patient with a combination of HIV    inhibitors; followed by-   (ii) the intermittent administration of a pharmaceutical composition    comprising an anti-virally effective amount of TMC278 or a    pharmaceutically acceptable acid-addition salt thereof, in    accordance with the present invention, wherein the composition is    administered at a time interval of at least one week.

The present invention also concerns a pharmaceutical composition asdescribed hereinbefore for use as a medicament in the treatment orprophylaxis of HIV infection.

In addition, the present invention concerns the use of a pharmaceuticalcomposition as described herein for the preparation of a medicament forthe prophylaxis or treatment of HIV infection.

The present invention further concerns a method of treating a subjectinfected with HIV, said method comprising the administration of atherapeutically effective amount of a pharmaceutical composition asdescribed herein.

As used herein, the word “substantially” does not exclude “completely”e.g. a composition which is “substantially free” from Y may becompletely free from Y.

Where necessary, the word “substantially” may be omitted from thedefinition of the invention. The term “about” in connection with anumerical value is meant to have its usual meaning in the context of thenumerical value. Where necessary the word “about” may be replaced by thenumerical value ±10%, or ±5%, or ±2%, or ±1%.

All documents cited herein are incorporated by reference in theirentirety.

The following examples are intended to illustrate the present inventionand should not be construed as limiting the invention thereto.

EXAMPLE 1 Preparation of Nanosuspensions

Glass bottles of 250 ml and ZrO₂ beads, used as the milling media, weresterilized in an autoclave. Five grams of drug substance were put intothe 250 ml glass bottle as well as a solution of 1.25 g Pluronic F108 in60 ml of water for injection. Three hundred grams of ZrO₂-beads with anaverage particle size of 500 μm were added. The bottle was placed on aroller mill. The suspension was micronized at 100 rpm during 72 hours.At the end of the milling process the concentrated nanosuspension wasremoved with a syringe and filled into vials. The resulting formulationis Formulation 1 in the following table. Determination of theconcentration was done by HPLC/UV. A dilution was made to a finalconcentration of 25 mg/ml of TMC278. The resulting suspension wasshielded from light.

Using similar procedures, Formulations 2, 3 and 4 were prepared. Thesewere titrated with sodium hydroxide 1N solution to a pH of about 7. Informulations 2, 3 and 4 the Lipoid™ EPG is solubilized in the Tween™ 80.

Ingredient Formula 1 Formula 2 Formula 3 Formula 4 TMC278   5 g 300 mg 300 mg  300 mg  Pluronic ™ F108 1.25 g — — — Tween ™ 80 — 75 mg 75 mg 75mg Lipoid ™ EPG — 9.375 mg   9.375 mg   9.375 mg   Glucose — 50 mg 50 mg50 mg NaH₂PO_(4.)1aq — —  2 mg  2 mg citric acid.1aq — — —  1 mg NaOH 1N — at pH 6.72 at pH 6.98 at pH 6.99

EXAMPLE 2 Kinetic Study

The present study demonstrates that an injectable formulation ofnanonised TMC278 or its HCl-salt results in stable blood plasma levelsduring a prolonged period of time. This study compares the plasmakinetics of TMC278 base and hydrochloric salt after single intramuscular(IM) or subcutaneous injection of a nanosuspension (Formula I of theprevious example) at 5 mg/kg in male beagle dogs.

Six healthy male beagle dogs with body weights ranging from 8 to 16 kgat the start of the study, were used. Each dog was identified by an eartattoo number. Two dogs were dosed intramuscularly (IM) in the left andright m. biceps femoris (treatment group A). Two dogs were dosed IM withTMC278.HCl (treatment group B). Two dogs were dosed subcutaneously (SC)in the left and right thoracal region. The injection volume was 2×0.1ml/kg in all treatment groups. A 20 G needle was used.

Blood samples of 3 ml were taken from the left jugular vein from alldogs on day 0 at 0 h (predose), 20 min, 1 h, 3 h, 8 h and 24 h post-doseand further on days 2, 3, 6, 8, 10, 13, 16, 20, 23, 27, 29, 36, 43, 50,57, 64, 71, 78, 85 and 92 at approximately 8 AM. Blood samples wereplaced on EDTA, EDTA Vacuette Greiner, Cat. No. 454086, GreinerLabortechnik N.V.). Within 2 h of blood sampling, samples werecentrifuged at room temperature at about 1900×g for 10 minutes to allowplasma separation. Plasma was immediately transferred into a second tubeand stored in the freezer within 2 hours after the start ofcentrifugation. Plasma samples were analysed individually for TMC278 bymeans of a validated LC-MS/MS-method.

TABLE 1 Individual and mean plasma concentrations and some basicpharmacokinetic parameters of TMC278 in male beagle dogs after single IMadministration of a nanosuspension of TMC278 at 5 mg/kg Treatment groupA Compound TMC278 Dose (mg/kg) 5 Adm. Route IM Dog No Time (h) 1726418186 Mean   0 (d0) <1.00 <1.00 <1.00   0.33 121 186 154   1 110 82.696.3   3 131 145 138   8 130 136 133  24 (d1) 150 120 135  48 (d2) 159132 146  72 (d3) 115 99.6 107  144 (d6) 86.2 91.9 89.1  192 (d8) 72.475.5 74.0  240 (d10) 56.7 62.5 59.6  312 (d13) 33.4 38.0 35.7  384 (d16)23.9 20.6 22.3  480 (d20) 20.5 16.6 18.6  648 (d27) 11.4 9.08 10.2  696(d29) 11.3 11.2 11.3  864 (d36) 7.33 6.44 6.89 1032 (d43) 5.19 5.18 5.191200 (d50) 3.40 3.25 3.33 1368 (d57) 3.00 3.00 3.00 1536 (d64) 2.84 2.442.64 1704 (d71) 2.48 1.84 2.16 1872 (d78) 1.79 1.45 1.62 2040 (d85) 1.991.61 1.80 2208 (d92) 1.56 1.25 1.41 Cmax (ng/ml) 159 186 173 Tmax (h) 480.33 24 AUC0-312 h (ng · h/ml) 27400 26600 27000 AUC0-696 h (ng · h/ml)34800 33000 33900 AUC0-2208 h (ng · h/ml) 40500 38200 39400

TABLE 2 Individual and mean plasma concentrations and some basicpharmacokinetic parameters of TMC278 in male beagle dogs after single IMadministration of a nanosuspension of TMC278.HCl at 5 mg (eq.)/kgTreatment group B Compound TMC278.HCl Dose (mg eq./kg) 5 Adm. Route IMDog No Time (h) 19072 19080 Mean   0 (d0) <1.00 <1.00 <1.00   0.33 4.424.68 4.55   1 7.80 7.19 7.50   3 14.7 16.3 15.5   8 32.2 27.1 29.7  24(d1) 50.1 69.8 60.0  48 (d2) 85.6 105 95.3  72 (d3) 47.5 69.5 58.5  144(d6) 48.3 62.3 55.3  192 (d8) 46.8 65.8 56.3  240 (d10) 55.7 82.2 69.0 312 (d13) 27.0 45.8 36.4  384 (d16) 17.0 31.9 24.5  480 (d20) 13.7 25.519.6  648 (d27) 7.91 14.4 11.2  696 (d29) 10.2 18.8 14.5  864 (d36) 6.1811.4 8.79 1032 (d43) 6.32 8.18 7.25 1200 (d50) 4.56 5.68 5.12 1368 (d57)4.73 5.08 4.91 1536 (d64) 4.47 4.43 4.45 1704 (d71) 3.38 3.90 3.64 1872(d78) 3.12 3.20 3.16 2040 (d85) 3.20 4.00 3.60 2208 (d92) 2.96 2.81 2.89Cmax (ng/ml) 85.6 105 95.3 Tmax (h) 48 48 48 AUC0-312 h (ng · h/ml)15000 20900 18000 AUC0-696 h (ng · h/ml) 20300 30500 25400 AUC0-2208 h(ng · h/ml) 27400 39900 33600

TABLE 3 Individual and mean plasma concentrations and some basicpharmacokinetic parameters of TMC278 in male beagle dogs after single SCadministration of a nanosuspension of TMC278 at 5 mg/kg Treatment groupC Compound TMC278 Dose (mg/kg) 5 Adm. Route SC Dog No Time (h) 1912919349 Mean   0 (d0) <1.00 <1.00 <1.00   0.33 <1.00 <1.00 <1.00   1 1.621.37 1.50   3 7.96 8.42 8.19   8 27.6 13.8 20.7  24 (d1) 15.7 28.5 22.1 48 (d2) 34.8 29.1 32.0  72 (d3) 26.1 30.6 28.4  144 (d6) 18.9 32.7 25.8 192 (d8) 17.7 23.0 20.4  240 (d10) 24.3 42.0 33.2  312 (d13) 21.7 38.830.3  384 (d16) 21.7 16.6 19.2  480 (d20) 29.8 21.2 25.5  648 (d27) 19.011.0 15.0  696 (d29) 21.0 10.5 15.8  864 (d36) 12.7 5.49 9.10 1032 (d43)5.22 6.03 5.63 1200 (d50) 6.37 3.40 4.89 1368 (d57) 4.78 2.52 3.65 1536(d64) 6.45 2.05 4.25 1704 (d71) 3.96 3.57 3.77 1872 (d78) 3.66 1.91 2.792040 (d85) 8.60 2.82 5.71 2208 (d92) 3.05 2.49 2.77 Cmax (ng/ml) 34.842.0 38.4 Tmax (h) 48 240 144 AUC0-312 h (ng · h/ml) 6910 9880 8400AUC0-696 h (ng · h/ml) 15900 16700 16300 AUC0-2208 h (ng · h/ml) 2640022400 24400

1. A pharmaceutical composition for administration by intramuscular orsubcutaneous injection, comprising a therapeutically effective amount ofTMC278, a salt, a stereoisomer or a stereoisomeric mixture thereof, inthe form of a suspension of micro- or nanoparticles comprising: (a)TMC278, a salt, a stereoisomer or a stereoisomeric mixture thereof, inmicro- or nanoparticle form, having a surface modifier adsorbed to thesurface thereof; and (b) a pharmaceutically acceptable aqueous carrier;wherein the TMC278 active ingredient is suspended.
 2. A compositionaccording to claim 1 wherein the TMC278 is present as the E-isomer ofthe base form.
 3. A composition according to claim 1, wherein thesurface modifier is selected from the group of poloxamers, α-tocopherylpolyethylene glycol succinates, polyoxyethylene sorbitan fatty acidesters, and salts of negatively charged phospholipids.
 4. A compositionaccording to claim 1, wherein the surface modifier is selected fromPluronic™ F108, Vitamin E TGPS, Tween™ 80, and Lipoid™ EPG
 5. Acomposition of claim 1, wherein the average effective particle size ofthe TMC278 micro- or nanoparticles is below about 50 μm, in particularbelow about 200 nm.
 6. A composition of claim 1, wherein the averageeffective particle size of the TMC278 micro- or nanoparticles is about130 nm.
 7. A composition according to claim 1, comprising by weightbased on the total volume of the composition: (a) from 3% to 50% (w/v),or from 10% to 40% (w/v), or from 10% to 30% (w/v), of TMC278; (b) from0.5% to 10%, or from 0.5% to 2% (w/v) of a wetting agent; (c) from 0% to10%, or from 0% to 5%, or from 0% to 2%, or from 0% to 1% of one or morebuffering agents; (d) from 0% to 10%, or from 0% to 6% (w/v) of aisotonizing agent (e) from 0% to 2% (w/v) preservatives; and (f) waterfor injection q.s. ad 100%.
 8. The use of a pharmaceutical compositionas defined in claim 1, for the manufacture of a medicament for thetreatment of HIV infection, or for the prevention of HIV infection in asubject at risk of being infected by HIV.
 9. The use of claim 8 whereinthe medicament is for the long-term treatment of HIV infection, or forthe long-term prevention of HIV infection in a subject at risk of beinginfected by HIV.
 10. The use according to claim 8 wherein the medicamentis for administration by intramuscular or subcutaneous injection;wherein the composition is administered intermittently at a timeinterval of one week to two years.
 11. The use according to claim 8wherein the pharmaceutical composition is administered at an interval ofat least one month to one year.
 12. The use according to claim 8,wherein the pharmaceutical composition is administered at a timeinterval that is in the range of one week to one month, or in the rangeof one month to three months, or in the range of three months to sixmonths, or in the range of six months to twelve months, or in the rangeof 12 months to 24 months.
 13. The use according to claim 8, wherein thepharmaceutical composition is administered once every two weeks, or onceevery month, or once every three months.
 14. A process for preparing apharmaceutical composition as defined in claim 1, comprising. (a)obtaining TMC278 in micronized form; (b) adding the micronized TMC278 toa liquid medium to form a premix/predispersion; and (c) subjecting thepremix to mechanical means in the presence of a grinding medium toreduce the average effective particle size.